Caffeine is commonly consumed during pregnancy. Currently, it is recommended that pregnant women reduce caffeine intake to less than 300 milligrams per day. Yet, because caffeine only causes malformations of the fetus at extremely high doses, low dose exposure to the drug has not been well studied. There should be concern over fetal exposure to caffeine because caffeine crosses both the placental and blood brain barriers to accumulate in the fetal brain. The proposed studies will investigate the effects of low dose (equivalent to 2-3 cups of coffee per day) chronic prenatal exposure to caffeine on fetal neurodevelopment. The first set of experiments will determine whether prenatal caffeine exposure alters the function and/or number of receptors/channels in the hippocampus during development. Calcium imaging techniques will be used to determine whether moderate prenatal caffeine exposure alters calcium regulation in hippocampal neurons. Also, Western blot analysis will be used to determine whether altered calcium transients are due to changes in the numbers of receptors/channels expressed. These studies are important because caffeine has indirect effects at many ion channels and receptors due to its anatagonism of adenosine receptors. The next set of experiments will determine whether prenatal caffeine exposure alters cell death and cell number in the developing hippocampus. If prenatal caffeine exposure enhances cell death due to altered receptor/channel function, there may be a loss of neurons in the hippocampus. TUNEL analysis will be performed to measure levels of cell death at various time points and immunohistochemistry will be used to estimate total neuron number in the hippocampus of caffeine exposed and control rats. Lastly, behavioral measures of learning and memory will be performed in juvenile rats to determine if there are long-lasting developmental consequences of prenatal caffeine exposure. These studies are relevant to public health because caffeine is commonly consumed during pregnancy and its use during pregnancy may have effects on fetal brain development and long-term effects on mental health. These changes in brain development may manifest as learning impairments that are not diagnosed until adolescence or adulthood. Thus, it is extremely important and relevant to determine whether low dose chronic exposure to caffeine has effects on the developing fetal brain.